LABSG
Journal of the American Association for Laboratory Animal Science
Volume 49, Number 5, September 2010
ORIGINAL RESEARCH
Reproduction
Schuler et al. Measurement of Fetal Biparietal Diameter in Owl Monkeys (Aotus nancymaae), pp. 560-563
Domain 3: Research; TT3.1. Biomethodology technique
Tertiary Species: Other Nonhuman Primates
SUMMARY: Owl monkeys are valuable animal models for malaria, ocular and other infectious diseases. Determining gestational age is important for breeding colonies and biparietal diameter can be used to determine gestational age. The BPD is the measurement of the diameter of the fetal head made when the outer calvarium is clearly visible. This technique has been used in squirrel monkeys and capuchins but never in owl monkeys. This report concluded that BPD can be used to predict gestational age of owl monkey fetuses and ultrasound can be used to predict fetal viability.
QUESTIONS:
1. What is the genus and species or the owl monkey?
2. Which of the following harbor cercopithicine herpesvirus 1?
a. Pan troglodytes
b. Macaca fascicularis
c. Aotus nancymaae
d. Saguinus Oedipus
3. What is the gestation of the owl monkey?
a. 143-146 days
b. 60-63 days
c. 132-135 days
d. 152-155 days
ANSWERS:
1. Aotus nancymaae
2. b
3. c
Husbandry
Arbona et al. Treatment and Eradication of Murine Fur Mites: I. Toxicologic Evaluation of Ivermectin-Compounded Feed, pp. 564-570
Task 1: Prevent, Diagnose, Control, and Treat Disease
Primary Species: Mouse
SUMMARY: Treatment of fur mites with ivermectin is discussed. Mice (free of rodent pathogens) were fed ivermectin-containing feed (12, 24, and 48 ppm) ad libitum. Control mice were fed an unmedicated diet. At experimental time points, mice were euthanized with CO2 and blood and tissues were collected. Ivermectin concentration was measured in plasma and skin. Ivermectin concentration was also measured in feed after processing and storing as a function of time. Differences in body weight and ivermectin levels in feed, plasma, and skin were compared by using the Kruskal-Wallis test when testing for overall differences among many groups. The Wilcoxon signed rank test was used to compare differences in paired groups.
The manufacturing process affected ivermectin concentrations with lower-than-expected concentrations in the pellets. Ivermectin was stable in the pellets for 6 mos post-mill date.
No significant difference was noted in body weight between groups. However, a transient difference was noted at 2 time-points but resolved over the course of the experiment. In all groups, plasma ivermectin levels plateaued at day 7-10. 9 days after ending treatment, plasma ivermectin was no longer detectable. Reproductive performance was not different among groups. However, pup mortality was significantly higher in the 24- and 48- ppm groups and was dose-dependent.
C57BL/6 mice seemed to be resistant to high doses of ivermectin. Only 1 of 144 mice exhibited clinical signs. The authors suggest that a dose of 1.3 mg/kg body weight for an 8-wk period is an acceptable regimen.
QUESTIONS:
1. What is the most common genus/species of fur mite in murine colonies?
2. From what class of drugs does ivermectin belong?
3. What route(s) can ivermectin be administered?
ANSWERS:
1. Myobia musculi and Myoptes musculinus
2. Avermectin
3. PO, SC, topical
Bennett et al. Multidimensional Cost-Benefit Analysis to Guide Evidence-Based Environmental Enrichment: Providing Bedding and Foraging Substrate to Pen-Housed Monkeys, pp. 571-577
Domain 4: Animal Care; Task 1 – Develop animal husbandry programs
Primary Species: Bonnett Macaque
SUMMARY: Environmental enrichment is a key component to modern animal care programs, and is intended to promote the physical and psychological well-being of laboratory animals. However, because of the multitude of variables involved in different scientific reports on environmental enrichment programs it is difficult to perform a cost-benefit analysis of different approaches to environmental enrichment. This study conducts a cost analysis on a previously demonstrated enhancement to a primate environment, e.g. bedding in pen-housed monkeys. Specifically this study attempts to capture all costs, including materials, labor and impact upon facility operation. The findings of the study show that contrary to initial impressions using bedding in a large indoor primate pen can be done effectively and can also reduce labor and materials costs while having a minimal impact on facility operation and animal care programs. Because soiled bedding was “spot-removed” and replaced with clean bedding labor costs were less than those associated with cleaning bare cages, and materials costs were reduced using less disinfectant solution. Furthermore simple solutions were found to prevent bedding clogging in drain pipes, the primary concern in regards to facility operation. Overall, this study shows that using bedding in primate pens can be a viable component of a primate environmental enrichment program.
QUESTIONS:
1. How can bedding promote species specific behavior in NHPs?
2. What are the major cost categories that should be considered when conducting a cost benefit analysis of environmental enrichment programs?
ANSWERS:
1. Bedding helps promote foraging and provides a more natural substrate for animals than do bare pens. Also reduces aggression, over grooming, and other abnormal behaviors.
2. Cost categories should include: Labor, materials and supplies (both one-time initial purchases and ongoing expenses) and facilities maintenance costs.
Pavek et al. Operation of an Air Filtration Device Results in Morbidity and Mortality in Growing Chickens (Gallus gallus domesticus), pp. 578-582
Tertiary Species: Chicken (Gallus gallus domesticus)
Domain 4: Animal Care; Task 3. Manage or provide indirect management/oversight of laboratory animal facilities; K11. Environmental causes of physiological alterations in animals and their effects on research (e.g., sound, light, temperature, humidity, housing systems)
Abstract: There are numerous reports demonstrating that changes in an animal's normal environment can be considered stressful and lead to negative consequences. This study was undertaken when results from studies on Marek's disease virus performed in newly acquired cubicles differed from results of experiments performed in a previous vivarium. This caused the authors to wonder if vivarium-specific environmental factors were the cause of the differences. In this study the authors prospectively evaluated the effect of an air filtration device ( AFD) on mortality and morbidity in a cohort of young chickens from a MDV research colony still in their growth phase.
Materials and Methods: All chickens in the study were from SPF P2a Leghorn breeding stock and hatched as a single group. At d1 chickens were randomly assigned without sexing to one of three groups: a control "Historical group" that was not exposed to the AFD, an AFD-off (negative control) group, and an AFD- on group. All cages were set-up alike with the only difference being whether the AFD was on or off; when "on" the AFD was "on" continuously for the 6 week duration of the study. All standard environmental parameters were monitored including temperature, relative humidity, noise, and light at time 0, 2,4 and 6 weeks. Room air particulate matter was measured at designated time points and averaged over a 30-second sample period for both nonfiltered aerosolized particulate and particulate capable of entering the respiratory airways. Body weights were measured at 2,4, and 6 weeks. Any birds showing any signs of ill health were euthanized and a body weight and gross necropsy were performed. All birds alive at the end of 6 weeks were euthanized and the degree of chronic stress was evaluated by obtaining adrenal, bursal, splenic, and left ventricular (cardiac) weights. Organ weights were normalized to correct for body weight differences by calculating organ:body weight ratios. Before euthanasia at six weeks, 2 pairs of sex-matched chickens from each of the upper, middle, and lower 25th weight percentiles for each study group were evaluated for aggressive vs. non-aggressive behaviors. All study parameters were evaluated by appropriate statistical measures.
Results:
• Growth Rate and Survival: The growth rate at all time points was significantly lower for the AFD-on group compared to the other two groups. Survival was lower in the AFD-on group as well.
• Behavior: There appeared to be very few behaviors displayed in any of the groups overall. The AFD-on group displayed the most behaviors for the all parameters, but did not achieve statistical significance.
• Post-mortem Findings: Both AFD groups demonstrated significant decreases in splenic and cardiac weight indices.
SUMMARY: The authors' conclusions based on the increased morbidity and mortality coupled with the decreased organ weights observed post-mortem in the AFD-on group of chickens who were exposed to the constantly running AFD units long term were that these findings were caused the ambient sound created by the AFD. They were not able to identify a mechanism by which the AFD caused the decreased weights and decreased survival of the chickens.
QUESTIONS:
1. Oxidative damage has been found to result from ____________ exposure to sound levels __________ in numerous species.
A. Acute, 45-85dB
B. Chronic , greater than 85dB
C. Chronic, 45-85 dB
D. None of the above.
2. The hearing frequency range of chickens is:
A. 4-16Hz
B. 1-5 kHz
C. 1-70kHz
D. 0.35-3.15kHz
3. Macrovibration aversion and physiologic effects have been shown to occur in fully grown chickens. True or False.
ANSWERS:
1. B. chronic, greater than 85db
2. D. 0.35-3.15kHz
3. True
Health Surveillance
Arbona et al. Treatment and Eradication of Murine Fur Mites: II. Diagnostic Considerations, pp. 583-587
SUMMARY: This is the second part to a three part series on the treatment and eradication of murine fur mites. There were essentially four parts to this project. 1. The authors wanted to determine the most effective fur mite diagnostic methodologies. 2. Three commercial laboratories were sent positive control slides in sets of slides containing diagnostic skin scrapings to assess accuracy of the laboratory. 3. The authors wanted to determine the time required for a mouse to shed its entire hair coat to help determine if finding eggs or casings months after treatment revealed treatment failure. 4. Lastly, the authors attempted to validate the efficacy of soiled bedding sentinel health monitoring programs by exposing mice to various concentrations of soiled bedding infested with mites.
Results:
1. The paper describes an outbreak in a specific facility, and thus the facilities methodology of cellophane tape was determined to be inferior. The group then turned to skin scrape from a number of sites (head, back and ventrum) and direct examination of the pelt for terminal examinations. Results showed in this study that skin scrape was much more effective in producing a true positive as compared to the pelt examination. The authors recommended the skin scrape method, and stated that samples should be taken from the animal’s head and back to avoid false-positive results.
2. Two of the three commercial laboratories failed to detect at least one of the positive slides submitted. As a result, the authors recommend the routine submission of known positive slides with diagnostic samples.
3. Three strains of white mice were dyed and evaluated monthly and photographed to monitor changes in coat color. The results revealed that more than 8 months is needed for a mouse to shed its entire hair coat, and the authors inferred that due to how tightly the eggs attach to hair shafts, eggs found months after the end of treatment could indicate a successful treatment and be caused by normal shedding patterns.
4. A pilot study was conducted to determine the efficacy of transmission of fur mites by soiled bedding transfer. Results indicated that only 2.5% of the bedding (1 of 40 cages) needs to be mite-infested to lead to a positive soiled bedding sentinel within 8 weeks.
QUESTIONS:
1. Which species of fur mites is reported to preferentially inhabit the forehead and cheeks?
2. Which species reportedly prefers the ventral abdomen and inguinal areas?
3. In this study, what percentage of animals tested positive with the pelt examination?
4. True or False: these authors implemented a 3 month post treatment testing moratorium prior to initiating follow up testing.
ANSWERS:
1. Myobia musculi
2. Myocoptes musculinus
3. Zero
4. True
Tavakkol et al. Resident Bacteria Flora in the Skin of C57BL/6 Mice Housed under SPF Conditions, pp. 588-591
Domain 1 - Prevent, Diagnose, Control and Treat Disease
Primary Species - Mouse
SUMMARY: The purpose of this study was to characterize the bacterial flora colonizing the skin of a particular strain of mice housed in an SPF colony at the University of Washington. These mice are used to study skin interaction with biomaterials with the goal of decreasing bacterial infections by restoring cutaneous barrier function. 30 total male C57BL/6 mice were used. 24 of the mice were euthanized after which punch biopsies were taken from the skin of the dorsal thorax. The biopsies were either from skin unexposed to antiseptics or from skin that was clipped, depilated, cleansed with providone-iodine and alcohol. The biopsies were then cultured for either aerobic or anaerobic bacteria. Cultured isolates were identified using various phenotypic testing. They were also processed for DNA extraction, PCR amplifications, and sequencing. Skin samples were taken from an additional six mice which were frozen and sent out for density gradient gel electrophoresis (DGGE) separation and DNA sequencing.
The number of colonies isolated from prepped skin was, as expected, significantly lower than from unprepped skin. The bacteria species identified by 16S DNA sequencing were as follows:
• S. saprophyticus (n = 19)
• S. aureus (n=17)
• S. lentus (n=16)
• S. cohnii (n=9)
• S. xylosus (n=9)
• Enterococcus faecalis (n=5)
• S. nepalensis (n=3)
• Stenotrophomonas maltophilia, Microbacteria testaceum, Aerococcus viridans, Bacillus licheniformis, B. anthracis, B. subtilis, B. circulans (n=1 each)
Sequencing of the frozen skin samples identified DNA from several of the bacterial species already cultured. Other bacteria identified that were not cultured include Porphyromomonas gingivalis, Flavobacteria symbiont, Burkholderia cepacia, Brevundimonas nasdae, and Microbacteriaceae bacterium.
The results show that staphylococci are the predominant bacterial species present in murine skin with coagulase -negative species outnumbering S. aureus. The results also show that for the purposes of identifying and quantifying bacterial flora, culture techniques alone are insufficient. Direct molecular analysis can identify species that do not normally grow in culture. Finally, the composition of skin flora in SPF mice is likely to vary between facilities due to a number of factors. Therefore, the results of this study are only applicable to the strain and facility from which the data was obtained.
QUESTIONS:
1. Which bacterial genus was the most commonly cultured from the mice SPF C57BL/6 used in this study?
a. Staphylococcus
b. Enterococcus
c. Bacillus
d. Streptococcus
2. T/F: Culture techniques alone are sufficient for quantifying and identifying the microflora from skin samples.
3. Which of the following Staphylococcus species was not isolated from the mice in this study?
a. S. aureus
b. S. saprophyticus
c. S. epidermidis
d. S. cohnii
ANSWERS:
1. A
2. F
3. C (S. epidermidis is the predominant bacterial species in human skin.) To be removed from the LABSG email list, send a message to:
Management
Reynolds et al. Noise in a Laboratory Animal Facility from the Human and Mouse Perspectives, pp. 592-597
Task3 K2; Task9 K1 & K2
Primary Species - Mus musculus
SUMMARY: Do mice hear what humans hear? Is sound within a vivarium perceived by mice like it is by humans? This study attempted to shed light on this particular topic. The authors focused on mice in ventilated cages and how much of the noise around them can be heard and to what extent. They compared the noise from the ventilated rack itself, an animal cage changing station, and construction jack hammer. Bottom line is that the noise from these devices is actually perceived to a lesser extent in mice than it is in human beings. They showed this by measuring sound pressure levels at various locations in the room and in the mouse cage and seeing where these measurements fell within both the human hearing range and the mouse hearing range. Because these particular objects produce noise of a low frequency, these noises reach well into the ambient sound levels perceivable by humans, but not into the ambient sound level of mice. Mouse hearing range is in the ultrasonic level and ranges from 1-100KHz. Human hearing range is down in the 20Hz - 20kHz range, just barely into that of the mouse.
Audiograms of mice show a much less sensitive hearing ability for mice at low frequencies, and then hearing becoming more sensitive as the frequency increases. Hearing is most sensitive for mice at about 16KHz, while that of humans is around 1-4 KHz. Knowing the frequency range in which an animal or human can hear helps to weight the measurement of the decibel range that an animal or human can hear. This is called "A-weighting". In other words, the A-weighting de-emphasizes decibels that are not within the most sensitive frequency range to determine what sounds can be heard and what sounds can't be heard. Some sounds are heard by humans at high decibel levels, but low frequencies.
This theory is how the authors of this paper determined which sounds were audible by humans and which were audible by mice. In this paper, although humans could hear the sound of the ventilated rack blower, the cage changing station, and the jack hammer, because the frequency of the sound was lower than the hearing range of mice, it is thought that the mice cannot hear these noises. The important point to remember is that although they may not hear the noise, the mice may be able to feel the vibrations, and it is the vibrations that may lead to detrimental effects in rodent studies.
QUESTIONS:
1. How is it that the noise of a jack hammer, which produces noise of a high decibel level, can be heard by humans but not mice?
2. What is the hearing range of a mouse in frequency?
3. What are the OSHA limits for decibel exposure for an employee without hearing protection averaged over an 8 hour work day?
4. What is the limit of decibel exposure over an 8 hour work day that then requires annual testing/training?
ANSWERS:
1. The jackhammer produces noise of a high decibel level, but a low frequency. This frequency is low enough and out of the range to where a mouse cannot accurately hear the noise.
2. Ultrasonic, 1-100KHz
3. 90Db
4. 85dB
Kanthaswamy et al. Population Genetic Statistics from Rhesus Macaques (Macaca mulatta) in Three Different Housing Configurations at the California National Primate Research Center, pp. 598-609
Domain 3: Research; Task 2: Advise and consult with investigators on matters related to their research
Primary Species: Rhesus macaques (Macaca mulatta)
SUMMARY: Population genetic diversity was evaluated at a National Primate Research Center colony of > 3000 rhesus macaques through evaluation of 15 short tandem repeats. Results were compared from three different housing settings, an indoor colony, outdoor corn cribs, and half-acre field cages. Current management activities to improve genetic diversity include selectively harvesting female macaques and cross-fostering infants among breeding cages. Results show that all three housing facilities have within-loci Hardy-Weinberg disequilibria, suggesting nonrandom associations among alleles at loci analyzed (decreased genetic diversity at the loci evaluated) as well as moderate to high levels of drift-driven differentiation among the housing facilities. The highest genetic diversity was observed in the indoor housing areas. This is not surprising as it is comprised of animals selected from the outdoor facilities and put on study and this population does not contribute to breeding (many are on terminal experiments and are not currently breeding). The investigators concluded that the current management methods being used at this facility are insufficient to prevent genetic subdivisions and evenly distribute genetic diversity throughout the colony. They also surmise that the inclusion of inbred or highly genetically homogenous animals with diminished allele diversity will likely compromise the potential for identifying allele-disease associations. Macaques of full-Chinese and full-Indian origins can exhibit profound phenotypic differences in physiologic and behavioral traits that are controlled by genetic mechanisms. Inclusion of macaques from different geographic regions or their hybrids in experimental research may confound interpretation of phenotypic differences due to inflation of the genetic contribution to phenotypic variances. To prevent this, the ancestry of rhesus macaques must be verifiable when animals with defined genetic or phenotypic characteristics are desired for research or breeding purposes. Excessive inbreeding can be avoided with the development of accurate marker-based pedigree records and sufficient numbers of unrelated animals available for breeding. In conclusion, balancing the sizes of matrilines, selective harvesting of females, and cross fostering of infants across housing units promotes genetic diversity and fosters gene flow, but may be inadequate as a sole strategy to maintain genetic diversity.
QUESTIONS:
1. List ways in which animal populations lose genetic diversity
2. T/F – Macaques of full-Chinese, full-Indian, or hybrid origin may be used interchangeably in all research studies.
ANSWERS:
1. Founder effect, intergenerational drift, nonrandom mating, genetic bottlenecks
2. False. Macaques of full-Chinese and full-Indian origins can exhibit profound phenotypic differences in physiologic and behavioral traits that are controlled by genetic mechanisms. Inclusion of macaques from different geographic regions or their hybrids in experimental research may confound interpretation of phenotypic differences due to inflation of the genetic contribution to phenotypic variances.
Experimental Use
Adamson et al. Assessment of Carprofen and Buprenorphine on Recovery of Mice after Surgical Removal of the Mammary Fat Pad, pp. 610-616
Domain 2, Tasks 1 and 2
Species: Mus musculus
SUMMARY: The authors of this study assessed the post operative recovery after the removal of the mammary fat pad in 4 week old female FVB mice. This procedure is generally considered minor, and analgesics are not often used. Food and water intake, body weight, wheel running activity and a video-recorded visual assessment were recorded for 4 days post procedure and compared with a presurgical baseline. Fecal corticosterone metabolite levels were measured at 12 and 24 hours post surgically. Groups were as follows: non surgical control, saline only, buprenorphine (0.2mg/kg SQ immediately post op and at 12 hours), carprofen (5 mg /kg SQ immediately post op), and a combination of the buprenorphine and carprofen groups.
Results: All surgical groups had a decrease in food consumption post surgery, and all groups returned to baseline levels by 48 hours. The buprenorphine and combination-treated mice had an acute drop in food intake at the 24 hour time point. Water intake was significantly lower at 24 hours for the combination treated group. The combination treated mice weighed significantly less at 24 hours post op. All groups had significantly higher weights by 96 hours post op. Wheel running activity was decreased for all groups at 24 hours post operatively compared with the non surgical controls. The buprenorphine group had an increased pain assessment score at 12 hours, and all groups had an increased pain assessment score at 24 hours. No groups were returned to baseline by 96 hours. Fecal corticosterone metabolite levels showed no significant differences across groups. The parameters used in their study did not indicate the chosen analgesic regimens improved recovery compared to the saline treated mice.
QUESTIONS:
1. What were the factors used to assess the pain score in the mice, and what postural movement indicative of pain was noted in all post surgical groups?
2. What treatment appeared to have the most negative effect on post operative recovery?
a. Saline
b. Buprenorphine
c. Carprofen
d. Combination of buprenorphine and carprofen
3. What important conclusions were made by the researchers regarding pain management in laboratory animals?
ANSWERS:
1. Hair coat, eyes, coordination and posture and overall condition were assessed in the scoring system. The abdominal stretch, and stretch and walk, were commonly noted in all post surgical groups.
2. b. Buprenorphine
3. Analgesic choice and the level of pain must be considered to help eliminate negative consequences on post operative recovery. Visual assessments of pain should be used carefully as some analgesics may have side effects that elevate the score. There is a need for further exploration of pain management dosing regimens and drugs in laboratory species.
Bourque et al. Comparison of Buprenorphine and Meloxicam for Postsurgical Analgesia in Rats: Effects on Body Weight, Locomotor Activity, and Hemodynamic Parameters, pp. 617-622
Primary Species: rat
Domain 2: Management of Pain and Distress; T1. Recognize pain and/or distress; T2. Minimize or eliminate pain and/or distress
SUMMARY: Buprenorphine is commonly administered for postoperative pain management in humans and animals, but there are complications associated with its use. Buprenorphine treatment in rodents has been shown to improve overall indications of pain and surgical stress, including postoperative food and water intake, changes in body weight, and ambient locomotor activity, compared with those of untreated controls. That said, unwanted effects associated with buprenorphine use in rodents include rebound hyperalgesia, variability in potency between sexes and strains of rodents, and issues with food intake (anorexia, pica). Furthermore, since buprenorphine treatment affects growth rate, feeding, and locomotor activity in naive rats, discriminating between behavioral cues caused by pain compared with those due to buprenorphine treatment per se is problematic. Finally, as an opioid analgesic, buprenorphine use is controlled in most jurisdictions, thereby complicating its availability. Meloxicam – a cyclooxygenase inhibitor that prevents synthesis of prostaglandin H2, a precursor to mediators that elicit pain and inflammation- is a potential alternative treatment for surgery in rats, but knowledge of its effects on recovery and cardiovascular parameters after surgery is incomplete. The objective of the present study was to compare buprenorphine (0.03 mg/ kg SC twice daily for 3 d) with meloxicam (2 mg/kg SC initial dose followed by 1 mg/kg SC once daily for 2 d) by assessing parameters relating to postsurgical recovery in 16 rats that underwent surgical implantation of radiotelemetric transducers. Implanting transducers in rats involves an abdominal incision of approximately 4 to 5 cm, visceral manipulation, implantation of a small catheter into the abdominal aorta, suturing the transducer to the inner muscular layer, and finally suturing the abdominal incision, making it a good model for testing post-operative analgesia. Meloxicam-treated rats had less severe behavioral (eating habits, locomotor activity) and physiologic (body weight loss, hemodynamic effects) manifestations than did buprenorphine-treated rats, either due to better analgesia, fewer pharmacologic side effects, or a combination thereof. Specifically, rats treated after surgery with buprenorphine showed greater reductions in body weight, food consumption, locomotor activity, and nighttime heart rates than did meloxicam-treated rats. Buprenorphine and meloxicam treatments both had stimulatory effects on mean arterial pressure and daytime heart rate measurements, although effects on nighttime mean arterial pressure were greater in the buprenorphine-treated rats. Authors conclude that these results suggest that meloxicam offers advantages over buprenorphine for use as a postoperative analgesic after laparotomy and radiotelemetric transducer implantation in rats.
QUESTIONS:
1. Which of the following is not an adverse effect reported with use of buprenorphine in rodents?
a. Rebound hyperalgesia
b. Variability in potency between sexes
c. Variability in potency between strains
d. Anorexia and/or pica
e. Renal toxicity
2. Which of the following statements is not supported in this study comparing post-operative use of buprenorphine and meloxicam in rats?
a. Meloxicam causes less reduction in body weight than buprenorphine
b. Meloxicam causes less reduction in food consumption than buprenorphine
c. Meloxicam causes less reduction in locomotor activity than buprenorphine
d. Meloxicam has less stimulatory effect on daytime heart rate measurements than buprenorphine
e. Meloxicam has less stimulatory effect on nighttime mean arterial pressure than buprenorphine
ANSWERS:
1. e. Renal toxicity
2. d. Meloxicam has less stimulatory effect on daytime heart rate measurements than buprenorphine
CASE REPORTS
Roble et al. A Comprehensive Laboratory Animal Facility Pandemic Response Plan, pp. 623-632
Domain 4: Animal Care
SUMMARY: As a result of the recent influenza outbreaks, national and local health organizations have encouraged employers to develop pandemic response plans to provide for protection of human life and assets. A disaster plan is also needed for animal facilities to provide guidance for the welfare of lab animals. The Rockefeller University Comparative Bioscience Center (RUCBC) established an Operation Continuity Plan to outline essential services of the animal research facility and develop a means to maintain those services in the case of a pandemic. The described plan is a 6-phase response to an event causing disruption of normal work patterns for 6 months or longer.
A University-wide pandemic work committee was established including members of human resources, lab safety and environmental health, finance, hospital, materials management, occupational health, security, plant operations, university residences, the vivarium, and RUCBC. An important element of the resulting pandemic plan was the classification of events as triggers for closing different parts of the University’s operations. The most severe response was cessation of the University’s activities and placement of the University into a state of “hibernation”. During a pandemic event, responsibility is reallocated from the President to an incident commander at an external agency. Additionally, a separate management hierarchy is established for the RUCBC. Support structures, such as housing, healthcare, etc., were identified that will help keep employees onsite. Financial and procurement procedures to ensure that key financial actions can be conducted were also evaluated.
Along with protecting human health, animal welfare must also be protected. Personnel with a wide variety of expertise within the animal facility were identified to help develop a special plan for the RUCBC that provided guidance beyond the University-wide plan. Plan contributors included veterinarians, managers, and staff involved in daily operations within the animal facility. Of note, IACUC approval should follow development of a plan. The RUCBC plan outlines a 2-tiered system of steps based on the WHO 8 level phase-alert classification system. Levels 5 and 6 correspond to declaration of a pandemic. The RUCBC plan also incorporates an internal severity index. Early level guidelines (level 0-1) incorporate education in disease transmission, hand sanitizer, and encouraging employees to stay home when sick. Levels 2-3 incorporate social distancing and PPE, such as N-95 respirators. Later pandemic stages include cessation of nonessential activities and alerting researchers to the step-by-step process of placing the University into hibernation mode. Hibernation mode is expected if employee absenteeism reaches 30-40% of the University workforce.
Since reduced staffing levels are expected during a severe pandemic, essential personnel are identified and expected to work on a rotating 2 week schedule. To facilitate this, personnel may be housed on campus during these times. Essential personnel are recruited on a voluntary basis with an incentive program including increased wages during the sequestered period, access to antiviral medication, and sabbatical time after the emergency. Volunteers are required to undergo physical and psychological evaluations. Documentation of these individuals is provided to local governments to facilitate travel and identification during disaster periods. Volunteers are assigned specific tasks, but are also cross-trained. During disaster periods, work shifts are 12 hr/ 7 d for two weeks. During this time, volunteers would remain on-site. Food provisions are kept in the form of ready-made meals. A 2-4 week supply is recommended.
Since food and bedding deliveries may be disrupted during a pandemic, larger quantities of food and bedding would need to be kept on-hand at one time. Advance discussion with suppliers enabled the RUCBC to establish a Memoranda of Understanding to facilitate bulk delivery of materials at the time of request. To extend the supply of bedding for small, rack-caged animals, the cage change frequency and the amount of bedding used per cage can be reduced. Cage change frequency may be reduced from once a week to twice a week, and with staff reductions beyond 30%, the frequency may need to be reduced even further. Frequent spot changes of dirty cages can extend the time between complete cage changes. Facilities that decrease the amount of bedding per cage should consider the type of bedding and absorbency, the cage size, and the number of animals per cage. Topping up of food supplies, rather than dumping and refilling hoppers can reduce the labor time and food required to feed animals. For large animals, facilities may choose to move animals from bedding to rubber matting to improve cage cleaning efficiency, and for nonhuman primates, nonperishable treats may be used instead of fresh fruits and vegetables which do not keep long in storage.
Due to the potential for food, bedding, and personnel shortages to have a negative impact on animal welfare, humane euthanasia of some animals should be considered. Investigator forethought is required to determine which rodent colonies to maintain. Under one plan, an investigator is given a quota of mouse cages that will be maintained and it is up to the investigator to determine how to distribute this quota. The RUCBC has also determined an order of which euthanasia will proceed, beginning with animals that require intensive care and ending with animals strains in production. Animals that are more specialized, such as internally derived strains, will be maintained as long as possible due to the difficulties in rederivation post pandemic. To minimize the psychological impact and reduce the time needed to euthanize large numbers of rodents, the RUCBC has purchased a CO2 euthanasia rack. In the event of animal euthanasia, the university has prepared two documents for communication. One document goes to internal sources, and the other goes to direct inquiries from outside sources. Accumulation of animal carcasses and waste must be considered, with municipal waste, cage waste, and biohazard waste considered individually. As with other critical services, facilities should arrange for critical and priority service guarantees from waste transportation and disposal services.
Long-term cessation of research projects may have lasting effects on research. To mitigate losses, investigators are encouraged to cryopreserve mice embryos and sperm, and important fish lines. Once the pandemic has subsided, employees are invited to return to work. All animal facilities that do not contain animals will be disinfected prior to animal reintroduction, and all essential personnel will be given time off.
QUESTIONS:
1. Why is cryopreservation recommended in the article?
2. Which of the following is NOT a major consideration when creating a pandemic response plan?
a. Recruitment of essential personnel
b. Food and bedding supplies
c. Animal euthanasia
d. Large scale utility outages
e. Disposal of animal carcasses
3. T/F. During an influenza outbreak, there is a potential for humans to pass the virus to animals in the research facility.
ANSWERS:
1. To mitigate research losses in the event of large scale animal euthanasia
2. d
3. T
Arbona et al. Treatment and Eradication of Murine Fur Mites: III. Treatment of a Large Mouse Colony with Ivermectin-Compounded Feed, pp. 633-637
Domain 1: Management of Spontaneous and Experimentally Induced Diseases and Conditions.
Primary Species: Mouse
SUMMARY: The investigators determined the efficacy of ivermectin-compounded feed against the fur mites Myobia musculi and Mycoptes musculinus. The authors designed a primary study to determine the necessary length of treatment regime which was followed by a large scale application to a 30,000 cage facility divided between 3 vivaria. The authors make a case for the ease of application of medicated feed versus other treatment strategies utilized for eradication of fur mites.
Evaluation of Treatment Efficacy: Female C57BL/6NCrl mice (n=32) were divided into 5 groups (A,B,C,D, and E). Groups A, B, and C were confirmed positive for both species of mites and treated with irradiated medicated chow (12ppm Ivermectin) for 1, 4, and 8 weeks respectively. Cages D & E were positive and negative controls. All treatment groups A, B, and C were negative for fur mites at the conclusion of the study and it was determined that a minimum treatment time should be exposure to medicated feed for at least a 7 day duration.
Facility-wide treatment: Mice received ivermectin-containing feed (12ppm @ at an estimated dose of 1.3mg/kg) for 8 consecutive weeks. Approximately 90% of the cages were treated with this approach. This was based on the acceptability of using the medicated feed with regards to ongoing research endeavors. Those animals that were not treated as above were treated with topical Selamectin solution at a final concentration of 4.5 mg/kg. The investigators were unable to detect fur mites during the follow up period 12 months post-treatment. Both sentinels and skin scraping from the colony were used to monitor post treatment. The authors reported only 1 investigative group that experience adverse effects from the treatment strategy. The projects involved genetically engineered mice that had received intracranial injections to induce CNS tumors. The animals died much earlier than expected. It was postulated that disruption of the blood brain barrier might account for the increased levels of ivermectin in the CNS.
QUESTIONS:
1. True/False Ivermectin administration has been associated with toxicity in some mouse strains and stocks.
2. How long is the life cycle for Myobia musculi and Mycoptes musculinus?
3. True/False Chronic ascariasis may provoke secondary amyloidosis due to long-standing dermatitis.
ANSWERS:
1. True
2. Myobia musculi = 23 days Mycoptes musculinus = 8-14 days
3. True
Schutt and Turner. Microchip-Associated Sarcoma in a Shrew (Suncus murinus), pp. 638-641
Domain 1: Management of Spontaneous and Experimentally-Induced Diseases and Conditions; T3: Diagnose disease or condition as appropriate; K5: anatomic pathology including pathogenesis of significant naturally occurring (e.g. tuberculosis) and experimentally induced (e.g. collagen-induced arthritis) diseases; typical gross and histopathologic lesions (e.g. age-related changes, or pathologic changes of adverse phenotypes associated with genetically modified rodents); and pertinent anatomic pathology techniques (e.g. Steiner's stain)
Tertiary Species: Other Rodents
SUMMARY
Background: House musk shrews (Suncus murinus) are members of the order Insectivora that are similar in size to domestic mice. They are used in research involving the emetic reflex and antiemetic pharmaceutical testing due to their ability to vomit in response to toxic stimuli. All animals in this colony were microchipped at 1 month of age because of their inherent research and breeding value. This particular animal was noted to have a rapidly growing, expansile mass between the scapulae at the site of microchip implantation at 16 months of age. The animal was humanely euthanized and submitted for necropsy.
Materials and Methods: The intrascapular mass was excised and submitted for histopathology. Marked gross splenomegaly was also noted at necropsy. Slides were stained with hematoxylin and eosin. Additional tumor immunohistochemistry staining for vimentin, pancytokeratin, alpha-smooth muscle actin, S100, desmin-specific enolase, and neuron-specific enolase was also performed.
Results: Hematoxylin and eosin staining revealed a population of interweaving, storiform spindle-shaped cells with numerous mitotic figures, compatible with a poorly-differentiated sarcoma. The neoplastic cells stained 95% positive for neuron-specific enolase, 5% positive for alpha-smooth muscle actin, and negative for all other immunohistochemical markers tested. Marked extramedullary hematopoeisis was noted in the spleen, liver, adrenal glands, and left kidney.
Discussion: The pathology findings are consistent with a fibroblastic or poorly-differentiated soft tissue sarcoma of non-neuromuscular origin, such as a fibrosarcoma. This is the first report of microchip-associated sarcoma in a house musk shrew. Microchips are commonly used to identify many species of laboratory animals, and microchip-associated neoplasms have also been reported in mice, rats, a dog, an Egyptian fruit bat, a degu, and a feathertail glider; all of these tumors have been malignant and mesenchymal in origin. A small amount of splenic hematopoeisis is normally present in mature shrews and supplements bone marrow hematopoiesis throughout life. The marked extramedullary hematopoiesis in this animal may be a response to the necrotic tissue core within the tumor.
QUESTIONS:
1. What is the genus and species of the house musk shrew?
a. Suncus murinus
b. Mus musculus
c. Rousettus aegyptiacus
d. Octogon degus
e. Acrobates pygmaeus
2. Microchip-associated neoplasms of laboratory animals are generally classified as:
a. Neuroendocrine and malignant
b. Epithelial and benign
c. Neuromuscular and benign
d. Mesenchymal and malignant
e. Round-cell and malignant
3. Vimentin is an immunohistochemical marker for which type of tissue?
a. Neural
b. Endocrine
c. Mesenchymal
d. Smooth muscle
e. Adipose
ANSWERS:
1. A
2. D
3. C
Hamir and Rupprecht. Pulmonary Idiopathic Alveolar Ossification in a Raccoon (Procyon lotor), pp. 642-643
SUMMARY: A ~ two year old male wild caught raccoon was treated experimentally with 4mg/kg lead acetate PO for 8 weeks. At the end of the treatment period the raccoon was euthanized, a complete necropsy was performed, and multiple tissues were submitted for histopathologic analysis. Bilateral thyroid adenomas, pancreatic islet cell amyloidosis, renal cortical infarcts, cystic adenomatous hyperplasia of the bladder uroepithelium, and cerebrovascular mineralization were noted and were reported to be incidental and previously documented in raccoons. Additionally ossified foci were noted in the alveolar tissue. Pulmonary ossification is rare and has not been previously reported in raccoons. Hypercalcemia, hyperphosphatemia, alkalosis, and lung injury are predisposing conditions that have been associated with pulmonary parenchymal ossification, but were not documented in this animal. The pulmonary ossification did not appear to be associated with the numerous incidental lesions re ported or the lead acetate treatment.
QUESTIONS:
1. Name the genus and species of the common raccoon.
2. What is a common histochemical stain for amyloid?
3. What modified histochemical stain was used in this study to identify intracytoplasmic and intranuclear lead inclusions?
ANSWERS:
1. Procyon lotor
2. Congo red
3. Ziehl -Neelsen
Asfaw and Sun. Presumed Mycobacteriosis in Laboratory Zebra Finch (Taeniopygia guttata), pp. 644-646
Domain 1, Task 3, K4
Tertiary Species – Other Birds [Zebra Finch (Taeniopygia guttata)]
SUMMARY: The most common agents of avian mycobacteriosis are Mycobacterium avium intracellulare (one of the group of bacteria known as MAC -Mycobacterium Avium Complex) and Mycobacterium genavense. MAC can cause disease in immunocompromised persons and is of zoonotic concern. Mycobacteria are saprophytic, aerobic and common in the soil. In this case study, a zebra finch was reported to be seen tossing its head back and forth and was eventually euthanized. Acid-fast (Ziehl-Neelsen) staining revealed rod shaped bacteria in macrophages of lung and duodenum, which led to a presumptive diagnosis of systemic mycobacteriosis. Deposits of black brown material often seen with pneumoconiosis were also present in some of the macrophages in the lungs. There are 2 common presentations of mycobacteriosis- the tubercular and atypical forms. The tubercular form appears as discrete granulomatous nodules seen in the later stages of the chronic disease process ( which was found in the liver of the Zebra finch). The atypical form is characterized by a conglomeration of histiocytes containing acid-fast bacilli, causing enlargement of organs (similar to the findings in the duodenum). In addition to the careful selection of mycobacteriosis free/pathogen free birds, effective cleaning with tuberculocidal properties as well as instituting an adequate quarantine program is warranted for preventing disease.
QUESTIONS:
1. Which form of presumed mycobacteriosis was found in the duodenum of the Zebra finch?
2. Which of the following is the scientific name of the Zebra finch?
a. Spinus cucullatus
b. Carduelis carduelis
c. Taeniopygia guttata
3. Which stain was used to identify the organisms presumed to be Mycobacterium spp?
ANSWERS:
1. Atypical form
2. C. Taeniopygia guttata
3. Acid-fast (Ziehl-Neelsen)
Daviau and Merton. Nonsurgical Repair of a Pseudoaneurysm in a Cynomolgus Macaque (Macaca fascicularis), pp. 647-651
Task 1: Prevent, diagnose, control and treat disease
Task 2: Prevent, alleviate and minimize pain and distress
Primary Species: Macaques (Macaca spp.)
SUMMARY: Pseudoaneurysm results when there is a leak of arterial blood into a defined space. In this case report, a pseudoaneurysm was diagnosed after presentation of a cyno for evaluation of an edematous and ecchymotic leg 1 week post femoral vein blood collection. The animal was on a neurotoxicity study receiving twice weekly saphenous vein injections of manganese sulfate, with chronic weekly blood collections. Pain and distress was not evident, but upon physical exam a large undefined mass in the femoral region was palpated. The animal was evaluated by Doppler ultrasound, and blood flow into the mass occurred during systole, and reversed during diastole, consistent with pseudoaneurysm, as well as a biphasic waveform in the femoral artery below the mass and pulsatile flow in femoral vein with the cardiac cycle that was consistent with an arteriovenous fistula between the femoral artery and vein. Ultrasound guided compression repair was recommended to avoid a significant risk of fatal hemorrhage with surgical repair. Induction of thrombosis of the mass with thrombin injections could not be performed due to the presence of the fistula, which presented a risk factor for seeding thrombin induced clots to the lung and CNS. The UGCR was performed by visualization and then compression of the neck of the pseudoaneurysm for 10 minutes with the ultrasound probe, followed by a second 10 minute session over the pseudoaneurysm to successfully induce coagulation and thrombosis. A third 10 minute session was used to close the arteriovenous fistula, and was confirmed with Doppler ultrasound. The animal remained anesthetized for the following 3 hours to permit the masses to solidify. Follow-up ultrasound confirmed successful stabilization and closure of the pseudoaneurysm, although the fistula recurred several months later.
QUESTIONS:
1. Pseudoaneurysm and formation of AV fistulas are common complications of femoral blood draws in NHP. T/F
2. What feature can be palpated to differentiate between a hematoma and pseudoaneurysm?
3. UGCR requires: a) an ultrasound probe b) a stent c) surgical access of the pseudoaneurysm d) Doppler verification of the absence of an AV fistula
ANSWERS:
1. False, this is rare, although the study does not report rates of occurrence.
2. A thrill can be palpated in a pseudoaneurysm as the blood flows with the cardiac cycle, a hematoma is static.
3. A
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